• Journal of Periodontal and Implant Science
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• v.53 no.5
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• pp.321-335
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• 2023

Purpose: The aim of this study was to investigate the efficacy of photo-crosslinked gelatin methacryloyl (GelMa) hydrogel containing calcium phosphate nanoparticles (CNp) when applying different fabrication methods for bone regeneration. Methods: Four circular defects were created in the calvaria of 10 rabbits. Each defect was randomly allocated to the following study groups: 1) the sham control group, 2) the GelMa group (defect filled with crosslinked GelMa hydrogel), 3) the CNp-GelMa group (GelMa hydrogel crosslinked with nanoparticles), and 4) the CNp+GelMa group (crosslinked GelMa loaded with nanoparticles). At 2, 4, and 8 weeks, samples were harvested, and histological and micro-computed tomography analyses were performed. Results: Histomorphometric analysis showed that the CNp-GelMa and CNp+GelMa groups at 2 weeks had significantly greater total augmented areas than the control group (P<0.05). The greatest new bone area was observed in the CNp-GelMa group, but without statistical significance (P>0.05). Crosslinked GelMa hydrogel with nanoparticles exhibited good biocompatibility with a minimal inflammatory reaction. Conclusions: There was no difference in the efficacy of bone regeneration according to the synthesized method of photo-crosslinked GelMa hydrogel with nanoparticles. However, these materials could remain within a bone defect up to 2 weeks and showed good biocompatibility with little inflammatory response. Further improvement in mechanical properties and resistance to enzymatic degradation would be needed for the clinical application.

• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.427-445
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• 2007

To improve osseointegration at the boneto-implant interface, several studies have been carried out to modify titanium surface. Variations in surface texture or microtopography may affect the cellular response to an implant. Osteoblast-like cells attach more readily to a rougher titanium surface, and synthesis of extracellular matrix and subsequent mineralization were found to be enhanced on rough or porous coated titanium. However, regarding the effect of roughened surface by physical and mechanical methods, most studies carried out on the reactions of cells to micrometric topography, little work has been performed on the reaction of cells to nanotopography. The purpose of this study was to examme the response of osteoblast-like cell cultured on blasted surfaces and alkali treated surfaces, and to evaluate the influence of surface texture or submicro-scaled surface topography on the cell attachment, cell proliferation and the gene expression of osteoblastic phenotype using ROS 17/2.8 cell lines. In scanning electron micrographs, the blasted, alkali treated and machined surfaces demonstrated microscopic differences in the surface topography. The specimens of alkali treatment had a submicro-scaled porous sur-face with pore size about 200 nm. The blasted surfaces showed irregularities in morphology with small(<10 ${\mu}m$) depression and indentation among flatter-appearing areas of various sizes. Based on profilometry, the blasted surfaces was significantly rougher than the machined and the alkali treated surfaces (p$TiO_2$) were observed on alkali treated surfaces, whereas not observed on machined and blasted surfaces. The attachment morphology of cells according to time was observed by the scanning electron microscope. After 1 hour incubation, the cells were in the process of adhesion and spreading on the prepared surfaces. After 3 hours, the cells on all prepared surfaces were further spreaded and flattened, however on the blasted and alkali treated surfaces, the cells exhibited slightly irregular shapes and some gaps or spaces were seen. After 24 hours incubation, most cells of the all groups had a flattened and polygonal shape, but the cells were more spreaded on the machined surfaces than the blasted and alkali treated surfaces. The MTT assay indicated the increase on machined, alkali treated and blasted surfaces according to time, and the alkali treated and blasted surfaces showed significantly increased in optical density comparing with machined surfaces at 1 day (p<0.01). Gene expression study showed that mRNA expression level of ${\alpha}\;1(I)$ collagen, alkaline phosphatase and osteopontin of the osteoblast-like cells showed a tendency to be higher on blasted and alkali treated surfaces than on the machined surfaces, although no siginificant difference in the mRNA expression level of ${\alpha}\;1(I)$ collagen, alkaline phosphatase and osteopontin was observed among all groups. In conclusion, we suggest that submicroscaled surfaces on osteoblast-like cell response do not over-ride the one of the surface with micro-scaled topography produced by blasting method, although the microscaled and submicro-scaled surfaces can accelerate osteogenic cell attachment and function compared with the machined surfaces.

• Journal of Periodontal and Implant Science
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• v.30 no.3
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• pp.661-676
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• 2000

Gingivitis and periodontitis are infectious diseases in that microorganisms are the primary extrinsic cause of the diseases. the occurrence of gingivitis has been associated clearly with the presence of microorganisms at the disease site, and the histologic nature of the tissue involved is indicative of an inflammatory response induced by microorganisms. additional evidence for the microbial etiology of periodontal disease is that numerous antimicrobial agents are effective in reducing plaque accumulation and periodontal diseases. the purpose of this article is to analyze the antimicrobial effects of Pulsatilla koreana. Well-dried Pulsatilla koreana purchased from herbs distributor was ground and extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH). we have then applied each solution to the bacteria samples(Bacteroides forsythus, Streptococcus mutans, Streptococcus sanguis, Porphylomonas gingivalis, Actinobacillus actinomycetemcomitans, Eikenella corrodens, Prevotella intermedia, Actinomyces viscosus, Prevotella nigrescens , Rothia dentocariosa, Fusobacterium nucleatum, Pseudomonas aeruginosa, Staphylococcus aureus) collected from several organizations. To conduct susceptibility test(Kirby-Bauer method), plate contained each periodontopathic bacteria is spread extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH) and to measure the minimum inhibition concentration(MIC) of the bacteria against the solutions to ultimately determine antimicrobial effects of the solutions, insert bacteria sample into $20{\mu\ell}/{m\ell}$, $10{\mu\ell}/{m\ell}$, $5{\mu\ell}/{m\ell}$, $2.5{\mu\ell}/{m\ell}$ of each solution and control group(not contained solution) 1. Solution extracted into methanol did not show clear zone against all bacteria samples. Only P.nigrescens, S. mutans and S. sanguis in solution extracted into ethylacetate, S. mutans and S. anguis in solutions extracted into chlorform and Butyl alcohol showed clear zone against all bacteria samples. Solution extracted into Butyl alcohol showed clear zone against 13 types of bacteria, excluding P. gingivalis. 2. In Solution extracted into methanol, the bacteria samples grew in the highest concentrated plate, showing minimal variation from control group. 3. In Solution extracted into Butyl alcohol, S. aureus, P. intermedia, E. corrodens, A. actinomycetemcomitans, B. forsythus, P. gingivalis et al. showed decreased growth in the highest concentrated plate. P. auruginosa, R. dentocariosa, A. viscosus, P. nigrescens, S. mutans et al. showed decreased growth at MIC $20{\mu\ell}/{m\ell}$ and S. sanguis showed decreased growth at MIC $10{\mu\ell}/{m\ell}$. 4. By analyzing the MIC level through considering the results from Kirby-Bauer method, Solution extracted into methanol did not reveal any antimicrobial effects and Solution extracted into Butyl alcohol showed the highest antimicrobial effects In conclusion, it can be used the extracts of Pulsatilla koreana as wide spectrum antimicrobial agent.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.439-443
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• 2006

Biocompatibility and tissue regenerating capacity are essential characteristics in the design of collagenous biomaterials for tissue engineering. Attachment of glycosaminoglycans to collagen may add to these characteristics by creating an appropriate micro-environment. In this study, porous type I collagen matrices were crosslinked using dehydrothermal treatment and 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide, in the presence and absence of chondroitin sulfate (CS). The scaffold like discs in 3 mm diameter were inserted into the intralamellar stromal pockets of rabbit cornea. In 8 weeks of follow up, clinical evaluation including corneal neovascularization, opacity and transparency of the graft scaffold was performed, and the inflammatory reaction and migration of corneal fibroblast were evaluated histologically. No inflammation, neovascularization and opacity in any of the implant were observed. CS increased the corneal fibroblast invasion and the transparency. It is concluded that the type I collagen sponge showed a biocompatibility in corneal stromal layer and addition of CS slightly improved the quality of the bioartificial corneal stromal layer. These results could be useful for the development of corneal substitutes.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.129-142
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• 2002

Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.

• Journal of Periodontal and Implant Science
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• v.35 no.2
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• pp.335-343
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• 2005

Inorganic bovine bone mineral has been widely researched as bone substitution materials in orthopedic and oral and maxillofacial application. OCS-B(NIBEC, Korea) is newly-developed inorganic bovine bone mineral. The aim of this study is to evaluate the safety and efficacy of bovine bone-derived bone graft material(OCS-B). Micro-structure of newly-developed inorganic bovine bone mineral(OCS-B) was analyzed by scanning electron microscope(SEM). Round cranial defects with eight mm diameter were made and filled with OCS-B in rabbits. OCS-B was inserted into femoral quadrant muscle in mouse. In scanning electron microscope, OCS-B was equal to natural hydroxyapatite. Rabbits were sacrificed at 2 weeks and 4 weeks after surgery and mice were sacrificed at 1 week and 2 weeks after surgery. Decalcified specimens were prepared and observed by microscope. In calvarial defects, osteoid and new bone were formed in the neighborhood of OCS-B at 2 weeks after surgery. And at 4 weeks after surgery osteoid and new bone bridge formed flourishingly. No inflammatory cells were seen on the surface of OCS-B at 1 week and 2 weeks in mouse experimental group. It is concluded that newly-developed inorganic bovine bone mineral(OCS-B) is a flourishing bone-forming material and biocompatible material.

• Design & Manufacturing
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• v.11 no.2
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• pp.51-56
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• 2017

Nitinol, a shape memory alloy (SMA), is manufactured from titanium and nickel and it used in various fields such as electrical applications, micro sensors. It is also recommended as a material in medical for implant because it has excellent organic compatibility. Nitinol is intended to be inserted into the human body, products require a high-quality surface and low residual stress. To overcome this problems, explore electrolyte polishing (EP) is being explored that may be appropriate for use with nitinol. EP is a particularly useful machining method because, as a non contact machining method, it produces neither machining heat nor internal stress in the machined materials. Sandpaper polishing is also useful machining method because, as a contact machining method, it can easily good surface roughness in the machined materials. The electrolyte polishing (EP) process has an effect of improving the surface roughness as well as the film polishing process, but has a characteristic that the residual stress is hardly generated because the work hardened layer is not formed on the processed surface. The sandpaper polishing process has the effect of improving the surface roughness but the residual stress remains in the surface. We experimented with three conditions of polishing process. First condition is the conventional polishing. Second condition is the electrochemical polishing(EP). And Last condition is a mixing process with the conventional polishing and the EP. Surface roughness and residual stress of the nitinol before a polishing process were $0.474{\mu}mRa$, -45.38MPa. Surface roughness and residual stress of the nitinol after mixing process of the conventional polishing and the EP were $1.071{\mu}mRa$, -143.157MPa. Surface roughness and residual stress of the nitinol after conventional polishing were $0.385{\mu}mRa$ and -205.15MPa. Surface roughness and residual stress of sandpaper and EP nitinol were $1.071{\mu}mRa$, -143.157MPa. The result shows that the EP process is a residual stress free process that eliminates the residual stress on the surface while eliminating the deformed layer remaining on the surface through composite surface machining rather than single surface machining. The EP process can be used for biomaterials such as nitinol and be applied to polishing of wafers and various fields.

• The Journal of Korean Academy of Prosthodontics
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• v.49 no.3
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• pp.245-253
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• 2011

Purpose: The aim of this in vitro study was to estimate surface characteristic after peptide coating and investigate biological response of human mesenchymal stem cell to anodized titanium discs coated with RGD peptide by physical adhesion and chemical fixation. Materials and methods: Fluorescence isothiocyanate (FITC) modified RGD-peptide was coated on the anodized titanium discs (diameter 12 mm, height 3 mm) using two methods. One was physical adhesion method and the other was chemical fixation method. Physical adhesion was performed by dip and dry procedure, chemical fixation was performed by covalent bond via silanization. In this study, human mesenchymal stem cell was used for experiments. The experiments consisted of surface characteristic evaluation after peptide coating, analysis about cell adhesion, proliferation, differentiation, and mineralization. Obtained data are statistically treated using Kruskal-Wallis test and Bonferroni test was performed as post hoc test (P=.05). Results: The evaluation of FE-SEM images revealed no diffenrence at micro-surfaces between each groups. Total coating dose was higher at physical adhesion experimental group than at chemical fixation experimental group. In cell adhesion and proliferation, RGD peptide coating did not show a statistical significance compared with control group (P>.05). In cell differentiation and mineralization, physical adhesion method displayed significantly increased levels compared with control group and chemical fixation method (P<.05). Conclusion: RGD peptide coating seems to enhance osseointegration by effects on the response of human mesenchymal stem cell. Especially physical adhesion method showed more effective than chemical fixation method on response of human mesenchymal stem cell.

• The Journal of Korean Academy of Prosthodontics
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• v.45 no.1
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• pp.85-97
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• 2007

Statement of problem: Recently, anodic oxidation of cp-titanium is a popular method for treatment of titanium implant surfaces. It is a relatively easy process, and the thickness, structure, composition, and the microstructure of the oxide layer can be variably modified. Moreover the biological properties of the oxide layer can be controlled. Purpose: In this study, the roughness, microstructure, crystal structure of the variously treated groups (current, voltage, frequency, electrolyte, thermal treatment) were evaluated. And the specimens were soaked in simulated body fluid (SBF) to evaluate the effects of the surface characteristics and the oxide layers on the bioactivity of the specimens which were directly related to bone formation and integration. Materials and methods: Surface treatments consisted of either anodization or anodization followed thermal treatment. Specimens were divided into seven groups, depending on their anodizing treatment conditions: constant current mode (350V for group 2), constant voltage mode (155V for group 3), 60 Hz pulse series (230V for group 4, 300V for group 5), and 1000 Hz pulse series (400V for group 6, 460V for group 7). Non-treated native surfaces were used as controls (group 1). In addition, for the purpose of evaluating the effects of thermal treatment, each group was heat treated by elevating the temperature by $5^{\circ}C$ per minute until $600^{\circ}C$ for 1 hour, and then bench cured. Using scanning electron microscope (SEM), porous oxide layers were observed on treated surfaces. The crystal structures and phases of titania were identified by thin-film x-ray diffractmeter (TF-XRD). Atomic force microscope (AFM) was used for roughness measurement (Sa, Sq). To evaluate bioactivity of modified titanium surfaces, each group was soaked in SBF for 168 hours (1 week), and then changed surface characteristics were analyzed by SEM and TF-XRD. Results: On basis of our findings, we concluded the following results. 1. Most groups showed morphologically porous structures. Except group 2, all groups showed fine to coarse convex structures, and the groups with superior quantity of oxide products showed superior morphology. 2. As a result of combined anodization and thermal treatment, there were no effects on composition of crystalline structure. But, heat treatment influenced the quantity of formation of the oxide products (rutile / anatase). 3. Roughness decreased in the order of groups 7,5,2,3,6,4,1 and there was statistical difference between group 7 and the others (p<0.05), but group 7 did not show any bioactivity within a week. 4. In groups that implanted ions (Ca/P) on the oxide layer through current and voltage control, showed superior morphology, and oxide products, but did not express any bioactivity within a week. 5. In group 3, the oxide layer was uniformly organized with rutile, with almost no titanium peak. And there were abnormally more [101] orientations of rutile crystalline structure, and bonelike apatite formation could be seen around these crystalline structures. Conclusion: As a result of control of various factors in anodization (current, voltage, frequency, electrolytes, thermal treatment), the surface morphology, micro-porosity, the 2nd phase formation, crystalline structure, thickness of the oxide layer could be modified. And even more, the bioactivity of the specimens in vitro could be induced. Thus anodic oxidation can be considered as an excellent surface treatment method that will able to not only control the physical properties but enhance the biological characteristics of the oxide layer. Furthermore, it is recommended in near future animal research to prove these results.